The US president spent four days in Ayrshire and Aberdeenshire in July visiting his golf courses, while the US vice president spent around four days in Ayrshire the following month.
In a letter to Chief Secretary to the Treasury James Murray, Scotland's Finance Secretary Shona Robison said the visits imposed "substantial operational and financial burdens on Scottish public services, particularly Police Scotland".
She said that the estimated additional cost of policing the presidential visit alone was around £20 million, reflecting peak daily deployments of more than 4,000 officers, while costs for the vice president's trip were around £6 million.
Ms Robison said that while the trips were framed as private visits by the president and vice president, they were "diplomatically significant".
She wrote: "Following your decision not to provide funding to Scotland for costs incurred in relation to the visit of President Donald Trump to Scotland in July 2025 and the subsequent visit of Vice President JD Vance, I am writing to you to request that you reconsider this decision and provide full reimbursement for the cost of the visits."
Ms Robison pointed out that the UK Government reimbursed the cost of Mr Trump's visit to Scotland in 2018.
Referring to the visits this summer, she wrote: "It is important to note that these visits were not initiated by the Scottish Government, nor were they part of devolved policy initiatives.
"Whilst I recognise this was not an official visit initiated by the UK Government, any visit by a sitting president and vice president will always constitute a high-profile event.
"There is clear previous precedent, where the UK Government has supported policing costs for visits to devolved nations by foreign dignitaries, including for the costs of the president's visit in 2018.
"Although the London element of the 2018 visit was for official engagements, the Scotland element was not."
She warned that failing to reimburse the costs would "not only strain devolved budgets but also set a troubling precedent for future high-profile visits".
The letter was sent last week.
Scotland's public finance minister Ivan McKee said: "We are clear that the UK Government should reimburse in full the significant costs of the working visits by President Trump and Vice President Vance.
"The visits were part of UK Government international relations, with the Prime Minister formally meeting the president in two locations in Scotland during his visit.
"It is completely unacceptable to expect the Scottish Government to foot the bill for what were clearly not private visits, as the UK Government is claiming."